Primary rat lacrimal cells undergo acinar-like morphogenesis on reconstituted basement membrane and express secretory component under androgen stimulation.
Single cells or small cell clusters, isolated from the rat lacrimal gland, were incubated on reconstituted basement membrane (matrigel) in a well-defined serum-free medium. During the first days of culture, cells reassociated and reorganized in structures resembling acini. These multicellular structures, maintained in culture for 2 weeks, consisted of well-polarized cuboidal cells surrounding a central lumen and exhibiting apically located microvilli. Myoepithelial cells were observed at the periphery of the acinar structures. Both in the native lacrimal and in the cultured aggregates, epithelial cells displayed strong immunoreactivity for cytokeratin 8, while myoepithelial cells were immunoreactive for vimentin and alpha-smooth muscle isoactin. These data indicate that the cultured aggregates closely mimic the in vivo architecture of lacrimal glands both by morphology and immunohistochemistry. We further demonstrated the presence of an intact androgen receptor and the ability of the cultured aggregates to respond to androgens with increased secretion of the secretory component. Comparable androgen responses were observed in lacrimal gland cultures of 5-week-old male and female rats. In conclusion, we report a morphologically and functionally differentiated culture system of primary rat lacrimal cells, in which androgen-regulated gene expression was observed. This culture model provides a unique experimental paradigm for studying the effects of hormones, cytokines, and growth factors on the morphogenesis, growth, and functional differentiation of lacrimal glands.
Vanaken
H
H
Division of Biochemistry, University of Leuven, Leuven, B-3000, Belgium.
Vercaeren
I
I
Claessens
F
F
De Vos
R
R
Dewolf-Peeters
C
C
Vaerman
J P
JP
Heyns
W
W
Rombauts
W
W
Peeters
B
B
D000199:Actins / Q000032:analysis; D000367:Age Factors; D000818:Animals; D001485:Basement Membrane; D018929:Cell Culture Techniques / Q000379:methods; D002467:Cell Nucleus / Q000737:chemistry; D002478:Cells, Cultured; D003094:Collagen; D004247:DNA / Q000096:biosynthesis; D004338:Drug Combinations; D004847:Epithelial Cells; D005260:Female; D007633:Keratins / Q000032:analysis; D007765:Lacrimal Apparatus / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D007797:Laminin; D008297:Male; D015741:Metribolone / Q000494:pharmacology*; D009919:Orchiectomy; D011509:Proteoglycans; D051381:Rats; D017208:Rats, Wistar; D011944:Receptors, Androgen / Q000032:analysis; D012635:Secretory Component / Q000032:analysis*; D045165:Testosterone Congeners / Q000494:pharmacology*; D014746:Vimentin / Q000032:analysis
D000199:Actins; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D011944:Receptors, Androgen; D012635:Secretory Component; D045165:Testosterone Congeners; D014746:Vimentin; C059485:matrigel; D015741:Metribolone; D007633:Keratins; D003094:Collagen; D004247:DNA